Project Abstract Alcoholism and anxiety frequently co-occur in humans;however, it has been difficult to find a single treatment which is effective against both conditions. Substantial evidence suggests that the motivational aspects of alcohol withdrawal [e.g., increased anxiety and anhedonia] referred to as negative affective states play an important role in the maintenance of excessive alcohol drinking, and may also be associated with relapse. Evidence also suggests a salient role for GABAergic mechanisms in regulating excessive alcohol drinking and the negative affective states associated with abstinence. The initial objective of the present proposal is to identify novel [unreadable]1 GABAA subtype-preferring ligands at the preclinical level that may serve as prototypes for further evaluation of clinical efficacy in treating both excessive alcohol drinking and the negative affective states associated with abstinence. To accomplish this, Aim 1A will employ our established pharmacophore/receptor model of BDZ binding sites to synthesize novel [unreadable]1 subtype-preferring ligands with reduced efficacies at diazepam sensitive [DS] subtypes [e.g., [unreadable]1-3]. Once the three agents [e.g., [unreadable]CCt, 3-PBC, WYS8] have been synthesized, Aim 1B will test the hypothesis that their chronic oral administration for 30 consecutive days can effectively attenuate excessive binge alcohol drinking in high alcohol drinking [HAD] rats. Aim 1C will test the hypothesis that a similar treatment will attenuate the negative affective states associated with abstinence. We hypothesize that chronic BDZ treatments will attenuate both binge dependence drinking and the negative affective states associated with abstinence. The second objective will be to identify select GABAA receptor subunits which may play a role in the regulation of excessive alcohol drinking and the negative affective states associated with abstinence. Aim 2A will test the hypothesis that inhibition of the [unreadable]1 receptor subunits within the ventral pallidum [VP] will lead to selective time-dependent reductions in binge alcohol responding, while Aim 2B will evaluate the hypothesis that their inhibition within the lateral/basolateral amygdala [BLA] will lead to time-dependent reductions in negative affective states. To downregulate the [unreadable]1 subunit, a novel siRNA sequence will be delivered into the VP and BLA by bilateral microinfusion using a herpes simplex virus-1 [HSV-1] amplicon vector. These studies should identify novel pharmacotherapies for further evaluation of clinical efficacy in treating comorbid alcoholism and anxiety at the preclinical level. In addition, they should shed light on the salient neuronal mechanisms in the regulation of the comorbid condition, which could be important in ultimately leading to a successful treatment for the comorbid condition.